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Table 1 Primers designed for scFv cloning and annealing temperatures.

From: Cloning approach and functional analysis of anti-intimin single-chain variable fragment (scFv)

Primers

Sequence

Annealing T°

VH Forward

5' GGATCC GTGCAGCTGCAGGAGTCTGG 3'

60°C

VH Reverse Linker

5' ACCGCCTCCACCGGAGACGGTGACCGTG 3'

76°C

VL Forward Linker

5' GGTGGCGGATCGGACATTGTGCTGACC 3'

78°C

VL Reverse

5' AAGCTT TTA GTTTGATTTCCAGCTTGGTGCC 3'

70°C

  1. Heavy chain forward primer contains the restriction site BamHI (bold). At the light chain reverse primer was inserted the restriction site HindIII (italic) and a stop codon (bold/italic)
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BMC Research Notes

ISSN: 1756-0500

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